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Clinical Chemistry Lecture > Analytical Methods and Instrumentations > Flashcards

Analytical Methods and Instrumentations Flashcards

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1
Q

Two primary considerations in
every colorimetric analysis:

A
  1. Quality of the color
  2. Intensity of the color
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2
Q

Special dye that changes color on the reactions

A

Chromogen

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3
Q

4 basic disciplines on Analytical Chemistry

A

Spectrometry
Luminescence
Electroanalytic Methods
Chromatography

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4
Q

Examples of Spectrometry

A

Spectrophotometry
Atomic Absorption
Mass Spectrometry

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5
Q

Examples of Luminescence

A

Fluorescence
Chemiluminescene
Nephelometry

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6
Q

Examples of Electroanalytic Methods

A

Electrophoresis
Potentiometry
Amperometry

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7
Q

Examples of Chromatography

A

Gas
Liquid
Thin-layer

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8
Q

2 Categories in Colorimetry

A

Visual Colorimetry
Photoelectric Colorimetry

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9
Q

Types of Photoelectric Colorimetry

A

Spectrophotometric Measurement
Photometric Measurement

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10
Q

Measurement of light intensity in a narrower wavelength.

A

Spectrophotometric Measurement

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11
Q

Measurement of light intensity at multiple wavelengths.

A

Photometric Measurement

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12
Q

Detection of light as a potential marker of concentration of analyte

A

Luminescence

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13
Q

Our of the 4 disciplines of Analytical Chemistry, what are the disciplines that uses light or radiant energy?

A

Spectrometry
Luminescence

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14
Q

Spectrophotometric uses what type of monochromators?

A

Prisms
Diffraction Gratings

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15
Q

Photometric uses what type of monochromators?

A

Filters

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16
Q

Measurements of Radiant Energy:

A

Emitted
Transmitted
Absorbed
Scattered
Reflected

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17
Q

Transmitted via electromagnetic waves that
are characterized by their frequency and Wavelength

A

Energy

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18
Q

The distance between two successive
peaks and it is expressed in terms of nanometer

A

Wavelength

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19
Q

Wavelength on visible spectrum?

A

400-700 nm

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20
Q

Wavelength on ultraviolet region?

A

<400 nm

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21
Q

Wavelength on infrared region?

A

> 700 nm

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22
Q

The relationship between wavelength and energy is described by _____

A

Planck’s Formula

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23
Q

Planck’s Formula:

A

E=hv

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24
Q

In Planck’s formula (E=hv), the E is _____ and the V is _____

A

E - is the energy of a photon in Joules or constant
V- frequency

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25
Q

Value of E in the Planck’’s formula:

A

6.626 x 10^27 erg sec

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26
Q

Types of Radiant Energy

A

Cosmic rays
Gamma rays
X-rays + Visible
Ultra-violet (UV)
Infrared (IR)
Radio, TV, microwave, etc.

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27
Q

The number of vibrations of wave motion per second

A

Frequency

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28
Q

The _____ the wave frequency, the longer the
wavelength.

A

Lower

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29
Q

The lower the wave frequency, the _____ the
wavelength.

A

Longer

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30
Q

The _____ the wavelength, the higher the
frequency and energy.

A

Shorter

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31
Q

The shorter the wavelength, the _____ the
frequency and energy.

A

Higher

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32
Q

Represents the wavelength in nanometers at peak transmittance.

A

Nominal Wavelength

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33
Q

Used to calibrate wavelength; quality control.

A

Didymium
Holmium Oxide Filters

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34
Q

Measurement of the light transmitted by a solution to determine the concentration of the light-absorbing substances in the solution.

A

Spectrophotometry

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35
Q

Ratio of the radiant energy transmitted, divided by the radiant energy incident on the sample.

A

Transmittance

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36
Q

Formula for Transmittance:

A

%T=(It/Io) x 100

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37
Q

In the formula for Transmittance (%T=(It/Io) x 100, the It is the _____ and the Io is the _____

A

It- radiant energy that is transmitted
Io - light that will pass through the exit slit towards the cuvette

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38
Q

The amount of light absorbed; proportional to the inverse log of transmittance.

A

Absorbance

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39
Q

Absorbance is mathematically derived from _____

A

%T

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40
Q

Formula for Absorbance:

A

A = 2 – log%T
A = -log%T
A = 1/log%T

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41
Q

States that the concentration of a substance is directly proportional to the amount of the light absorbed or inversely proportional to the logarithm of transmitted light.

A

Beer’s Law orBeer-Lambert’s Law

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42
Q

Formula for Beer’s Law orBeer-Lambert’s Law:

A

A=abc

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43
Q

In the formula of Beer’s Law or Beer-Lambert’s Law (A=abc), this is identified as the absorbance

A

A

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44
Q

In the formula of Beer’s Law or Beer-Lambert’s Law (A=abc), this is identified as the molar absorptivity

A

a

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45
Q

In the formula of Beer’s Law or Beer-Lambert’s Law (A=abc), this is identified as the length of light through the solution

A

b

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46
Q

In the formula of Beer’s Law or Beer-Lambert’s Law (A=abc), this is identified as the concentration of absorbing molecules/solution

A

c

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47
Q

It is the simplest type of absorption spectrometer designed to make one measurement at a time at one specified wavelength.

A

Single Beam Spectrophotometer

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48
Q

An Instrument that splits the monochromatic light into two components - one beam passes through the sample, and the other through a reference solution or blank. The additional beam corrects for variation in light source
intensity.

A

Double-beam Spectrophotometer

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49
Q

2 Types of Double-beam Spectrophotometer:

A

Double-beam in Space
Double-beam in Time

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50
Q

A type of Double-beam Spectrophotometer that has 2 photodetectors, for the sample
beam and reference beam.

A

Double-beam in Space

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51
Q

A type of Double-beam Spectrophotometer that has one photodetector and alternately
passes the monochromatic light through the sample cuvet and the reference cuvet using a chopper or rotating sector mirror.

A

Double-beam in Time

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52
Q

7 basic components of single or doublebeam configuration spectrophotometer:

A
  1. Stable source of radiant energy
  2. Filter that isolates a specific region of the electromagnetic spectrum
  3. Cuvets
  4. Sample holder
  5. Radiation detector
  6. Signal processor and
  7. Readout device
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53
Q

It provides polychromatic light and must generate sufficient radiant energy or power to measure the analyte of interest.

A

Light/Radiant Energy

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54
Q

To give accurate absorbance measurements throughout its absorbance range, its response to change in light intensity must be _____

A

Linear

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55
Q

2 Types of Light/Radiant Source:

A

Continuum Source
Line Source

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56
Q

A light source that emits radiation that changes in intensity; widely used in the laboratory.

A

Continuum Source

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57
Q

Examples of Continuum Source:

A

Tungsten
Deuterium
Xenon Lamps

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58
Q

Commonly used light source in the visible and near infrared region

A

Tungsten Light Bulbs

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59
Q

Routinely used light source to provide UV radiation in analytic spectrometers.

A

Deuterium Lamps

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60
Q

Light source that produces a continuous source of radiation, which covers both the UV and the visible range.

A

Xenon Discharge Lamp

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61
Q

Type of light source that emits limited radiation and wavelength.

A

Line Source

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62
Q

Examples of Line Source:

A

Mercury and sodium vapor lamps in
spectrophotometers
Hollow cathode lamp

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63
Q

LASER stands for _____

A

Light Amplification by Stimulated Emission of Radiation

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64
Q

It minimizes unwanted or stray light and prevents the entrance of scattered light into the monochromator system.

A

Entrance Slit

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65
Q

Refers to any wavelengths outside the band
transmitted by the monochromator; it does not originate from the polychromatic light source; it causes absorbance error.

A

Stray Light

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66
Q

The most common cause of loss of linearity at
high-analyte concentration.

A

Stray Light

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67
Q

Factors for choosing a light source:

A
  1. Range
  2. Spectral distribution within the range
  3. The source of radiant production
  4. Stability of the radiant energy and
  5. Temperature
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68
Q

It isolates specific or individual wavelength of light.

A

Monochromator

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69
Q

Alternatives for Tungsten Bulb:

A

Mercury arc (visible and UV)
Deuterium lamp (165nm) - UV
Hydrogen lamp - UV
Xenon lamp - UV
Merst glower -IR
Globar (silicone carbide) - IR

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70
Q

These are wedge-shaped pieces of glass, quartz or sodium chloride.

A

Prisms

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71
Q

Mercury Arc uses what type of radiant energy?

A

Visible
Ultraviolet

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72
Q

Deuterium Lamp uses what type of radiant energy?

A

Ultraviolet

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73
Q

Hydrogen Lamp uses what type of radiant energy?

A

Ultraviolet

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74
Q

Xenon Lamp uses what type of radiant energy?

A

Ultraviolet

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75
Q

Merst Glower uses what type of radiant energy?

A

Infrared

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76
Q

Globar (Silicone Carbide) uses what type of radiant energy?

A

Infrared

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77
Q

These are the most commonly used monochromators; better resolution than prism. These are made by cutting grooves (parallel grooves) or slits into an aluminized surface of a flat piece of crown glass.

A

Diffraction Gratings

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78
Q

Principle of Diffraction Gratings:

A
  • Wavelengths are bent as they pass a sharp corner.
  • Produces linear spectrum and therefore maintaining a constant and pass which is simple.
  • Can be used in the regions of spectrum where light energy is absorbed by glass prism.
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79
Q

These are simple, least expensive, not precise but useful monochromators. These are made by placing semi-transparent silver films on both sides of a dielectric such as magnesium fluoride.

A

Filters

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80
Q

Filters are made by placing semi-transparent silver films on both sides of a dielectric such as _____

A

Magnesium fluoride

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81
Q

Filters produce monochromatic light based on the principle of _____.

A

Constructive Interference

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82
Q

Constructive Interference principle:

A

Light waves enter one side of the filter and are reflected at the second surface.

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83
Q

Other term for Cuvet:

A

Absorption Cell
Analytical Cell
Sample Cell

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84
Q

Holds the solution whose concentration is to be measured.

A

Cuvet

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85
Q

Enumerate the Kinds of Cuvets:

A

Alumina Silica Glass
Quartz/Plastic
Borosilicate Glass
Soft Glass

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86
Q

A most commonly used cuvet.

A

Alumina Silica Glass

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87
Q

A cuvet used for measurement of solution
requiring visible and ultraviolet spectra

A

Quartz/Plastic

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88
Q

A cuvet that is used in acidic solutions.

A

Borosilicate Glass

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89
Q

A cuvet that is used for alkaline or base solutions.

A

Soft Glass

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90
Q

Silica cuvettes transmit light effectively at _____

A

Wavelengths 220mm

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91
Q

The path length of cuvets is _____

A

1cm

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92
Q

It contains cathode and anode enclosed in a glass case; it has a photosensitive material that gives off electron when light energy strikes it; requires an external voltage for operation.

A

Phototube

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93
Q

It detects and converts transmitted light into
photoelectric energy.

A

Photodetector

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94
Q

It detects the amount of light that passes through the sample in the cuvet.

A

Photodetector

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95
Q

It is the most commonly used detector that measures visible and UV regions.

A

Photomultiplier Tube (PMT)

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96
Q

It is the simplest detector; least expensive; temperature-sensitive.

A

Barrier Layer Cell

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97
Q

Other term for Barrier Layer Cell:

A

Barrier Layer Cell
Photocell
Photovoltaic Cell

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98
Q

Barrier Layer Cell is used in _____ with a wide bandpass.

A

Filter Photometers

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99
Q

Barrier Layer Cell is a basic phototransducer that is used for detecting and measuring radiation in _____

A

Visible Region

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100
Q

Type of Photodetector that is not as sensitive as PMT but with excellent linearity.

A

Photodiode

101
Q

Photodetector that measures light at a multitude of wavelengths - detects less
amount of light.

A

Photodiode

102
Q

Photodetector that has a lower dynamic range and higher noise compared to Photo Multiplier Tube.

A

Photodiode

103
Q

Photodetector that is most useful as a simultaneous multichannel detector.

A

Photodiode

104
Q

It displays output of the detection system

A

Meter or Read-out Device

105
Q

Examples of Meter or Read-out Device:

A

Galvanometer
Ammeter
LED Display

106
Q

The amount of light absorbed at a particular wavelength depends on molecular and
ion types present and may vary with:

A

Concentration
pH
Temperature

107
Q

Turbidity readings on a spectrophotometer are greater in the _____ than in _____ of the spectrum.

A

Blue Region
Red Region

108
Q

It measures the light emitted by a single atom burned in a flame.

A

Flame Emission Photometry

109
Q

Principle of Flame Emission Photometry:

A

Excitation of electrons from lower to higher energy state.

110
Q

Light Source in Flame Emission Photometry:

A

Flame

111
Q

Method used in Flame Emission Photometry:

A

Indirect Internal Standard Method

112
Q

Internal Standard in Flame Emission Photometry:

A

Lithium/Cesium

113
Q

In Flame Emission Photometry, this indicated changes in the fuel reading of the instrument.

A

Flickering Light

114
Q

Flame Emission Photometry is used for the measurement of excited ions such as:

A

Sodium
Potassium

115
Q

Purpose of Flame in FES:

A

Breaks the chemical bond to produce atoms. Source of energy absorbed by the atoms to enter an excited state.

116
Q

Breaks up the solution into finer droplets so that the atom will absorb heat energy from the flame and get excited.

A

Atomizer or Burner

117
Q

Means the blank contains serum but without the reagent to complete the assay.

A

Blanking Technique

118
Q

Corrects absorbance caused by the color of the reagents - the absorbance of reagents is automatically subtracted from each of unknown reading.

A

Reagent Blank

119
Q

Measures absorbance of the sample and reagent in the absence of the end product, and corrects the measurement for optical interference absorbing the wavelength of measurement.

A

Sample Blank

120
Q

_____ interfere mainly by increasing light blocked turbidity.

A

Lipids

121
Q

Methods or Procedures that is performed to correct for artifactual absorbance readings.

A

“Blanking” Procedures
Dual-wavelength Methods

122
Q

Interference Filters as Monochromators:

A

Na Filter
K Filter
Lithium

123
Q

Na Filter transmits what color of light at what wavelength?

A

Yellow Light
589 nm

124
Q

K Filter transmits what color of light at what wavelength?

A

Violet Light
767 nm

125
Q

Lithium transmits what color of light at what wavelength?

A

Red Light
761 nm

126
Q

Preferred internal standard; also acts as a radiation buffer in FES.

A

Lithium

127
Q

Reasons why Lithium is preferred:

A
  • Its emission characteristics are similar to those of Na+ and K+
  • Normally present as a trace element in human tissues and does not present interferences in the determination.
128
Q

Purpose of Lithium as a Quality Control in FES

A

To achieve stability where there is fluctuations caused by changes in fuel of air pressure which affects flame temperature and rate of sample aspiration.

129
Q

Used for measuring abundant large particles (proteins) and bacterial suspensions.

A

Turbidimetry

130
Q

Principle of Turbidimetry:

A

It determines the amount of light blocked (reduction of light) by a particulate matter in a turbid solution.

131
Q

Turbidimetry depends on:

A

Specimen Concentration
Particle Size

132
Q

Solutions requiring quantitation by turbidimetry are measured using:

A

Visible Photometers
Visible Spectrophotometers

133
Q

3 Sections in the Laboratory that utilizes Turbidimetry: (provide its uses in each sections)

A
  1. Clinical Chemistry - measure proteins
  2. Bacteriology - determination of bacterial growth
  3. Hematology - detect clot formation
134
Q

It measures the light absorbed by atoms dissociated by heat. Principle: Element is not excited by merely dissociated from its chemical bond and place in an unionized, unexcited, ground state

A

Atomic Absorption Spectrophotometry (AAS)

135
Q

Light source in Atomic Absorption Spectrophotometry (AAS):

A

Hollow-cathode Lamp

136
Q

Interferences in in Atomic Absorption Spectrophotometry (AAS):

A

Chemical
Matrix (differences in viscosity)
Ionization

137
Q

Atomic Absorption Spectrophotometry (AAS) is used for measurement of unexcited trace metals such as:

A

Calcium
Magnesium

138
Q

An atomizer such as _____ or _____ is used to convert ions to atoms while a _____ is used to modulate the light source.

A

Nebulizer
Graphite Furnance
Chopper

139
Q

What is added to samples to form stable complexes with phosphate.

A

Lanthanum
Stronium Chloride

140
Q

This is used for measuring the amount of antigen-antibody complexes (proteins)

A

Nephelometry

141
Q

Principle of Nephelometry:

A

It determines the amount of scattered light by a particulate matter suspended in a turbid solution

142
Q

In Nephelometry, light scattering depends on:

A

Wavelength
Particle Size

143
Q

Most antigen-antibody complexes have a diameter of _____ and the wavelengths used are _____, thus light is scattered forward.

A

250-1500nm
320-650m

144
Q

Detector used in Nephelometry:

A

Photomultiplier Tube

145
Q

Components of Nephelometer:

A

Light Source (Mercury Arc Lamp, Tungsten-Filament Lamp, Light Emitting Diode
LASER)
Collimator
Monochromator
Sample Cuvete
Stray Light Trap
Photodetector

146
Q

Principle of Volumetric:

A

The unknown sample is made to react with a
known solution in the presence of an indicator.

147
Q

Schales and Schales Method is used for _____

A

Chloride Test

148
Q

EDTA Titration Method is used for _____

A

Calcium Test

149
Q

Is the migration of charged particles in an electric field. It separates proteins on the basis of their electric charge and densities.

A

Electrophoresis

150
Q

Components of Electrophoresis:

A

Electrical power
Support Medium
Buffer
Sample
Detector Buffer: Barbital (pH 8.6)

151
Q

Has a net charge that can be either
positive or negative depending on pH conditions

A

Amphoteric

152
Q

The movement of buffer ions and solvent relative to the fixed support

A

Electroendosmosis/Endosmosis

153
Q

The migration of small charged ions

A

Iontophoresis

154
Q

The migration of charged macromolecules

A

Zone Electrophoresis

155
Q

Factors Affecting Rate of Migration:

A
  1. Net electric charge of the molecule
  2. Size and charge of the molecules
  3. Electric field strength
  4. Nature of the supporting medium
  5. Temperature of operation
156
Q

A Supporting Media that separates by molecular size

A

Cellulose Acetate

157
Q

A Supporting Media that separates by electrical charge; it does not bind protein.

A

Agarose Gel

158
Q

A Supporting Media that separates on the basis of charge and molecular size; separates proteins into 20 fractions; used to study isoenzymes.

A

Polyacrylamide Gel

159
Q

Amido black is used to visualize _____

A

Protein

160
Q

Ponceau s is used to visualize _____

A

Protein

161
Q

Oil Red O is used to visualize _____

A

Fats

162
Q

Sundan Black is used to visualize _____

A

Fats

163
Q

Fat Red 7B is used to visualize _____

A

Fats

164
Q

Coomassie Blue is used to visualize _____

A

CSF Protein

165
Q

A stain that is very sensitive even to nanogram quantities of proteins

A

Gold/Silver Stain

166
Q

It measures the absorbance of stain - concentration of the dye and protein fraction; it scans and quantitates electrophoretic pattern.

A

Densitometry

167
Q

It separates molecules by migration through a pH gradient; it is ideal for separating proteins of identical sizes but with different net charges.

A

Isoelectric Focusing

168
Q

pH gradient is created by adding _____ to the anodic area of the electrolyte cell and adding _____ to the cathode area.

A

Acid
Base

169
Q

Supporting Media for Isoelectric Focusing:

A

Agarose Gel
Polyacrylamide Gel
Cellulose Acetate

170
Q

Advantages of for Isoelectric Focusing:

A
  1. The ability to resolve mixture of proteins.
  2. To detect isoenzymes of ACP, CK and ALP in serum.
  3. To identify genetic variants of proteins such as alpha-1-antityrpsin.
  4. To detect CSF oligoclonal banding.
171
Q

In this method, sample molecules are separated by electro-osmotic flow (EOF); it utilizes nanoliter quantities of specimens.

A

Capillary Electrophoresis

172
Q

Capillary Electrophoresis utilizes _____ quantities of specimens.

A

Nanoliter

173
Q

Uses of Capillary Electrophoresis:

A

Separation
Quantitation and determination of molecular weights of proteins and peptides
Analysis of Protein products
Analysis of organic and inorganic substances and drugs

174
Q

It involves separation of soluble components in a solution by specific differences in physical-chemical characteristics of the different constituents.

A

Chromatography

175
Q

Bases of Separation:

A
  1. Rate of Diffusion
  2. Solubility of the solute
  3. Nature of the solvent
  4. Sample volatility/solubility
  5. Distribution between 2 liquid phases
  6. Molecular Size (molecular sieving)
  7. Hydrophobicity of the molecule
  8. Ionic attraction
  9. Differential distribution between two immiscible liquids
  10. Selective separation of substances
  11. Differences in adsorption and desorption of solutes
176
Q

2 Forms of Chromatography:

A

Planar
Column

177
Q

It is used for fractionation of sugar and amino acid.

A

Paper Chromatography

178
Q

Sorbent for Paper Chromatography

A

Whatman Paper

179
Q

It is a semiquantitative drug screening test.

A

Thin Layer Chromatography (TLC)

180
Q

What samples can be used for Thin Layer Chromatography (TLC)?

A

Blood
Urine
Gastric Fluid

181
Q

Sorbent for Thin Layer Chromatography (TLC)

A

Thin plastic plates impregnated with a layer of silica gel or alumina.

182
Q

It is used for separation of steroids, barbiturates, blood, alcohol and lipids; useful for compounds that are naturally volatile or can be easily converted into a volatile form.

A

Gas Chromatography (GC)

183
Q

In Gas Chromatography (GC), the samples are introduced into the GC column using a _____ or _____

A

Hypodermic Syringe
Automated Sampler

184
Q

Separation occurs based on differences in absorption at the solid phase surfaces.

A

Gas Solid Chromatography (GSC)

185
Q

Separation occurs by differences in solute partitioning between the gaseous mobile phase and the liquid stationary phase.

A

Gas Liquid Chromatography (GLC)

186
Q

Detector used for Gas Liquid Chromatography (GLC)

A

Flame Ionization

187
Q

It is based on the fragmentation and ionization of molecules using a suitable source of energy; it can also detect structural information and determination of molecular weight.

A

Mass Spectroscopy (MS)

188
Q

It is the gold standard for drug testing.

A

Gas Chromatography-Mass Spectroscopy (GC-MS)

189
Q

Gas Chromatography-Mass Spectroscopy (GC-MS) can also be used for:

A

Xenobiotics
Anabolic Steroids
Pesticides

190
Q

Can detect 20 inborn errors of metabolism from a single blood spot.

A

Tandem Mass Spectroscopy (MS/MS)

191
Q

It is based on the distribution of solutes between a liquid mobile phase and a stationary phase.

A

Liquid Chromatography

192
Q

The most widely used liquid chromatography.

A

High Performance Liquid Chromatography (HPLC)

193
Q

It uses pressure for fast separations, controlled temperature, inline detectors and gradient elution technique.

A

High Performance Liquid Chromatography (HPLC)

194
Q

Uses of High Performance Liquid Chromatography (HPLC):

A
  • Fractionation of drugs, hormones, lipids, carbohydrates and proteins
  • Separation and quantitation of various
    hemoglobins associated with specific diseases (e.g., thalassemia)
  • Rapid HbA1c test (within 5 minutes)
195
Q

In reverse phase HPLC, the _____ is more polar than the _____

A

Mobile Phase
Stationary Phase

196
Q

It is for detecting nonvolatile substances in body fluids.

A

Liquid Chromatography-Mass Spectroscopy (LC-MS)

197
Q

It is utilized to confirm positive results from screening of elicited drugs - it is a complementary method to GC-MS.

A

Liquid Chromatography-Mass Spectroscopy (LC-MS)

198
Q

Used in therapeutic drug monitoring, toxicology and studies of drug metabolites.

A

Liquid Chromatography-Mass Spectroscopy (LC-MS)

199
Q

Interface in Liquid Chromatography-Mass Spectroscopy (LC-MS):

A

Electrospray (ES)
Atmospheric Pressure Chemical Ionization (APCI)

200
Q

_____ is used in HPLC and GC methods to
compensate for variation in extraction.

A

Internal Standard

201
Q

The mechanism in this type of chromatography is the exchange of sample ions and mobile-phase ions with the charged group of the stationary phase.

A

Ion Exchange Chromatography

202
Q

Used for separation of amino acids, proteins and nucleic acids.

A

Ion Exchange Chromatography

203
Q

Separation of nucleic acids and proteins depends primarily on:

A

Charge
Ionic Charge Density

204
Q

Separation of compounds is based on their partition between a liquid mobile phase and a liquid stationary phase coated on a solid support.

A

Partition Chromatography (Liquid-Liquid
Chromatography)

205
Q

Used for separation of therapeutic drugs and their metabolites.

A

Partition Chromatography (Liquid-Liquid
Chromatography)

206
Q

It uses immobilized biochemical ligands as the
stationary phase to separate a few solutes from other unretained solutes.

A

Affinity Chromatography

207
Q

This type of separation uses the so-called lock-and-key binding that is widely present in biologic systems.

A

Affinity Chromatography

208
Q

Used for separation of lipoproteins, carbohydrates and glycated hemoglobins; antibodies.

A

Affinity Chromatography

209
Q

Separation is based on the differences (competition) between the adsorption and desorption of solutes at the surface of a solid particle.

A

Adsorption Chromatography (LiquidSolid Chromatography)

210
Q

In Adsorption Chromatography (LiquidSolid Chromatography), the compounds are adsorbed to a solid support such as:

A

Silica
Alumina

211
Q

It measures the amount of light intensity present over a zero background.

A

Fluorometry/Molecular Luminescence

212
Q

Principle of Fluorometry/Molecular Luminescence:

A

It determines the amount of light emitted by a molecule after excitation by electromagnetic radiation.

213
Q

Light detector for Fluorometry/Molecular Luminescence:

A

Photomultiplier Tube or Phototube

214
Q

Use of Fluorometry/Molecular Luminescence:

A

Porphyrins
Magnesium
Calcium
Catecholamines

215
Q

How many monochromators does Fluorometry/Molecular Luminescence use?

A

2 monochromators

  • The wavelength that is best absorbed by the solution to be measured is selected by the primary filter; the incident light is prevented from striking the photodetector by the secondary filter.
216
Q

Fluorometry/Molecular Luminescence is affected by quenching such as:

A

pH and temperature changes
Chemical contaminants
UV light change

217
Q

Light Source in Fluorometry/Molecular Luminescence:

A

Mercury arc lamp
Xenon lamp

218
Q

Emission of light is created from a chemical or
electrochemical reaction, and not from absorption of electromagnetic energy.

A

Chemiluminescence

219
Q

Principle of Chemiluminescence:

A

The chemical reaction yields an electronically
excited compound that emits light as it is ground state, or that transfers its energy to another compound, which then produces emission.

220
Q

Use of Chemiluminescence:

A

Immunoassays

221
Q

Photodetector in Chemiluminescence:

A

Photomultiplier tube (luminometer)

222
Q

It is the measurement of the osmolality of an aqueous solution such as serum, plasma, or urine.

A

Osmometry

223
Q

Principle of Osmometry:

A

It is based on measuring changes in the colligative properties of solutions that occur owing to variations in particle concentration.

224
Q

Osmotic Particles:

A

Glucose
Urea nitrogen
Sodium

225
Q

Colligative properties of the solution:

A

Osmotic pressure
Boiling point
Freezing point
Vapor pressure

226
Q

As the osmolality of a solution increases the following reactions occur: osmotic pressure _____; boiling point is _____; freezing point is _____; and the vapor pressure is _____.

A

Increases
Elevated
Depressed
Depressed

227
Q

The most commonly used method for measuring the changes in colligative properties of a solution.

A

Freezing-point Depression Osmometry

228
Q

A 1.0 mOsm/kg solution has a freezing point depression of _____ when compared with pure solvent (usually water).

A

0.00186 degree Celsius

229
Q

Blood plasma, with an osmolality of about _____, has a freezing point of about _____.

A

285 mOsm/kg
-0.53 degree Celsius

230
Q

The measurement of current or voltage
generated by the activity of a specific ion.

A

Electrochemistry Techniques

231
Q

It is the measurement of electrical potential due to the activity of free ions - change in voltage indicates activity of each analyte.

A

Potentiometry

232
Q

The measurement of differences in voltage (potential) at a constant current

A

Potentiometry

233
Q

Reference electrodes for Potentiometry:

A

Calomel
Silver-silver Chloride

234
Q

Use of Potentiometry:

A

pH test
pCO2 test

235
Q

It is an electrochemical transducer capable of responding to one given ion.

A

Ion Selective Electrode (ISE)

236
Q

ISE analyzers measure the _____ dissolved in the fluid phase of the sample in _____.

A

Electrolyte
mmol/L of plasma water

237
Q

2 types of Ion Selective Electrode (ISE):

A

1.Direct ISE (without sample dilution)
2.Indirect ISE (with sample dilution)

238
Q

Components of an ISE Membrane:

A

Glass aluminum silicate (sodium), valinomycin gel (potassium) organic liquid membrane ion exchangers (calcium and lithium) gas and enzyme electrodes

239
Q

Causes of Malfunctions of ISE:

A

Defective ISE membrane, buildup of counter voltages from liquid junction potentials at the salt bridge and buildup of proteins at the electrodes

240
Q

It is the measurement of the amount of
electricity (in coulombs) at a fixed potential.

A

Coulometry

241
Q

It is an electrochemical titration in which the
titrant is electrochemically generated and the
end point is detected by amperometry.

A

Coulometry

242
Q

Uses of Coulometry:

A

Chloride test in CSF, serum, and sweat

243
Q

Interference in Coulometry:

A

Bromide
Cyanide
Cysteine

244
Q

It is the measurement of the current flow
produced by an oxidation-reaction

A

Amperometry

245
Q

Use of Amperometry:

A

pO2
Glucose
Chloride
Peroxidase Determination

246
Q

It is the measurement of differences in current at a constant voltage.

A

Polarography

247
Q

The measurement of current after which a
potential is applied to an electrochemical cell.

A

Voltammetry

248
Q

Type of Voltammetry test used for lead and iron.

A

Anodic stripping voltammetry

Clinical Chemistry Lecture (3 decks)

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